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, 2019, 9845497
eCollection

The Human Amniotic Mesenchymal Stem Cells (hAMSCs) Improve the Implant Osseointegration and Bone Regeneration in Maxillary Sinus Floor Elevation in Rabbits

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The Human Amniotic Mesenchymal Stem Cells (hAMSCs) Improve the Implant Osseointegration and Bone Regeneration in Maxillary Sinus Floor Elevation in Rabbits

Lu Yin et al. Stem Cells Int.

Abstract

Insufficient bone height in the posterior maxilla is a challenging problem in dental implantation. Bio-Oss, though routinely used in maxillary sinus floor elevation (MSFE), is not osteoinductive. Human amniotic mesenchymal cells (hAMSCs) isolated from placental tissues have potential for multidifferentiation and immunomodulatory properties and can be easily obtained without the need for invasive procedures and without ethical concerns. This is the first study to use hAMSCs to improve implant osseointegration and bone regeneration after MSFE. Human AMSCs were loaded into a fibrin gel and injected into rabbit MSFE models. The rabbits were assigned to four groups (n = 3 per group), i.e., the control group, the hAMSC group, the Bio-Oss group, and the hAMSC/Bio-Oss group. The animals were sacrificed at postsurgery for four and twelve weeks and evaluated by histology and immunohistochemistry. Bone volume, bone volume/tissue volume, bone-to-implant contact ratio, and vessel-like structures in the hAMSC/Bio-Oss group were significantly better than those in other groups in the peri-implant and augmented areas. Immunofluorescence staining showed that alkaline phosphatase (ALP) activities of two hAMSC groups were higher than those of the other two groups. Sequential fluorescent labeling was performed in all of the 12-week groups. Observations showed that hAMSCs accelerated mineralized deposition rates on implant surfaces and in bone-augmented areas. These data demonstrated that hAMSCs could enhance implant osseointegration and bone regeneration after MSFE and might be used to optimize dental implantation in the future.

Conflict of interest statement

The authors have no conflicts of interest to disclose.

Figures

Figure 1
Figure 1
Schematic diagram of rabbit maxillary sinus floor elevation. (a) Implant and filling materials into the sinus: fibrin gel, human amniotic mesenchymal stem cells (hAMSCs), and Bio-Oss (Bio-Oss®, Geistlich Biomaterials, Wolhusen, Switzerland). (b) Four experimental groups. (c) Surgical procedure.
Figure 2
Figure 2
Characterization of human amniotic mesenchymal stem cells (hAMSCs). (a) Primary passage (P0) of hAMSCs. (b–e) Negative controls in eac h flow cytometric histogram. Human AMSCs were positive for CD105 (b), CD29 (b) and CD90 (c), and negative for CD34 (d) and HLA-DR (e). (f) hAMSCs at personal passage three (P3). (G-I) hAMSCs induced by osteogenic (g), adipogenic (h), and chondrogenic (i) media were stained by Alizarin Red S, Oil Red O, and Toluidine Blue solution, respectively.
Figure 3
Figure 3
Van Gieson's picro fuchsin (VG) staining. (a–h) VG staining of four groups in the 4- and 12-week groups. All of number 3 were partially enlarged views of number 2. All of numbers 4 and 5 were the partially enlarged views of number 1. (i–n) The amount of bone volume, bone volume/tissue volume, and percent of bone-to-implant contact ratio assessed at the same time points (i–k) and at various time points (l–n). BO: Bio-Oss; red dotted mark range: new bone; p < 0.05; ∗∗p < 0.01.
Figure 4
Figure 4
Immunohistochemical staining of α-smooth muscle actin (α-SMA). (a–h) Four groups in the peri-implant and augmented areas at four and 12 weeks. (i–l) Relative vessel-like structure at four and 12 weeks. BO: Bio-Oss; NB: new bone; red arrow: blood vessel-like structure. Scale bar = 200 μm. p < 0.05; ∗∗p < 0.01).
Figure 5
Figure 5
Immunofluorescence of alkaline phosphatase (ALP) and hematoxylin-eosin (HE) staining. (a–h) Four groups in the peri-implant and augmented areas at four and 12 weeks. (i–l) The positive ALP area at four and 12 weeks. DAPI: 4′,6-diamidino-2-phenylindole; BO: Bio-Oss; NB: new bone. Scale bar = 250 μm. p < 0.05; ∗∗p < 0.01.
Figure 6
Figure 6
Polychrome sequential fluorescent labeling. (a, b) calcein (CA), Alizarin Red (AL), and merged images in the peri-implant and augmented areas at different time points. Scale bar = 400 μm. (c, d) Relative area of fluorochrome-stained bone in the peri-implant and augmented area. p < 0.05; ∗∗p < 0.01.

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